Chemical Constituents Extraction And Antitumor Activity Of Dracocephalum Peregrinum Linn.

Dracocephalum peregrinum Linn.is a herbaceous plant of the genus Dracocephalum in the Labiatae family.It is a traditional Kazakh Chinese herbal medicine used to treat colds and liver diseases.At present,only few chemical and pharmacological studies were reported on D.peregrinum at home and abroad.Therefore,this paper has conducted a systematic study on the chemical composition and anti-tumor activity of D.peregrinum.?:Isolation and extraction of monomer compounds from D.peregrinumIn the paper,a systemic study of the chemical composition of the 70%ethanol reflux extract of D.peregrinum was carried out,using repeated silica gelcolumn chromatography,Sephadex LH-20 column chromatography,Semipreparative high performance liquid chromatography,ODS column chromatography,macroporous resin column chromatography and recrystallization.A total of 44 compounds were extracted and isolated from the whole plant of D.peregrinum,42 compounds were identified by Co-TLC,mass spectrumincluding,¹H nuclearmagnetic resonance spectroscopy and¹³C nuclearmagnetic resonance spectroscopy,including 13flavonoids and flavonoid glycosides:Luteolin(1),Kaempferol(2),Quercetin(3),Acacetin(4),Diosmetin(5),3,5-dihydroxy-6,7,3',4'-tetramethoxyflavone(6),Hesperetin(7),Alpinetin(8),Apigenin(9),Naringenin(10),Isoquercitrin(11),Isorhamnetin(12),Rutin(13);Six terpenoids:Oleanolic acid(14),Lupeol(15),Betulinic acid(16),Betulin(17),Ursolic acid(18),Corosolic acid(19);17 benzoic acid and phenylpropionic acid compounds:Ferulic acid(20),Phydroxycinnamic acid(21),3-Hydroxybenzoic acid(22),Vanillin(23),Syringate(24),Gallic acid(25),Chlorogenic acid(26),Caffeic acid(27),Methylcaffeate(28),dihydrocaffeic acid(29),Rosmarinic acid(30),Protocatechuic acid(31),Vanillic acid(32),Phenyl-?-D-glucoside(33),Arbutin(34),5-caffeoylquinic acid(35),4-dicaffeoylquinic acid(36);Four steroids:Stigmasterol(37),Stigmast-5-en-3?,7?-diol(38),?-Sitosterol(39),Daucosterol(40),2aliphatic compounds:Docosane(41),Linolenic acid(42).Among them,compound 6-8,10,12-13,19,24-25,29-30,32,34-36,38,42,17 compounds were isolated from D.peregrinum for the first time.?:Research on the anti-tumor activity of chemical constituents of D.peregrinumMTT assay was used to detect the effects of 42 monomer compounds isolated and purified from D.peregrinum and its 70%ethanol extract on A549,Hep G2,He La cells and L02,H8 two normal cells.The results show that 5(Diosmetin),13(Rutin),16(Betulinic acid),18(Ursolicacid),19(Ursolic acid)and 25(Gallic acid)significant inhibitory effect on the viability of three tumor cells;1(Luteoli)and 12(Isorhamnetin)inhibit the activity of Hep G2 cells;14(Oleanolic acid)inhibit the activity of He La cells.The IC50 value of 18 on tumor cells was less than 10?g/m L,showing good anti-tumor activity,but it also had significant inhibitory effect on normal cells.The IC50 values of 5,13,16,19 and 25 were higher on normal human hepatocytes than on tumor cells,the results prompt that 5,13,16,19 and 25 have good anti-tumor activity and drug safety.?:Study on the anti-glioma activity and its mechanism of diosmetinIn the paper,the anti-glioma cell activity and the potential mechanism of diosmetin,a flavonoid from D.peregrinum were studied.Firstly,MTT assay was used to detect the effect of diosmetin on the activity of U251 cells in glioma,the experimental results showed that the activity of cells in the U251 cell groups treated with different concentrations of diosmetin decreased,and the activity of cells decreased in turn with the increase of diosmetin concentration.Then,the effect of diosmetin on U251 cell apoptosis was studied by flow cytometry and western blot,the experimental results showed that diosmetin could induce U251 cell apoptosis by down-regulating Bcl-2 and up-regulating Bax and cleaved caspase-3 levels,and the apotosis rate increased significantly with the increase of diosmetin concentration.Finally,the effects of diosmetin on the migration and invasion of U251 cells in glioma were studied by using cell-scratching,transwell,western blot,real-time quantitative PCR and immunofluorescence staining.The experimental results showed that diosmetin possibly inhibit the migration and invasion of glioma cells by inhibiting TGF-?signaling pathway and activating E-cadherin expression.According to mouse tumor model experiments,diosmetin can inhibit the growth of glioma invivo.In the paper,the chemical constituents and antitumor activities of D.peregrinum were systematically studied,which laid the experimental foundation for the development and utilization of D.peregrinum.